Glucocorticoid sensitivity depends on expression levels of glucocorticoid receptors in canine neoplastic mast cells

Glucocorticoid (GC) administration with or without other chemotherapeutic reagents is a commonly used option in the treatment of mast cell malignancies. However, the responsiveness of mast cell tumors to GC treatment varies in individuals, and the regulatory mechanisms determining the GC sensitivity of malignant mast cells remain unclear. Since the expression of the GC receptor (GR) has been reported to be associated with GC sensitivity in human neoplastic lymphocytes, we attempted to investigate the relationship between GR levels and GC sensitivity by using neoplastic mast cells derived from canine mast cell tumors (MCTs). To elucidate the regulatory mechanisms involved in GC responsiveness, we analyzed various canine MCT cell lines and tissue samples from dogs with MCT. While the proliferation of canine MCT cells was suppressed by the addition of GC to the culture, we found that MCT cells derived from humans and rodents, as well as canine lymphoma cells, responded poorly to GC. However, there were also some variations in responsiveness to GC treatment among canine MCT cell lines used in this study. Using real-time polymerase chain reaction and Western blot analysis, we elucidated the relationship between GR expression and responsiveness to GC in canine MCT cells. Furthermore, to assess the involvement of GR expression in GC sensitivity in vivo, clinical investigations were conducted on dogs with cutaneous MCT. Written informed consent was obtained from owners, and the affected dogs were treated with prednisolone (0.5-2.0 mg kg(-1)day(-1), administered orally) 1 or 2 weeks prior to the surgical removal of the tumors. Tumor volume was measured according to WHO criteria both before and after prednisolone treatment, and the GC sensitivity of each MCT was determined on the basis of the reduction in tumor volume. Of the 15 dogs with MCT, 11 responded to treatment with prednisolone completely or partially, whereas 4 dogs showed no response. Examination of clinical samples obtained by surgical removal revealed that GR expression levels were significantly lower in GC-resistant MCT tissues than in GC-sensitive MCT tissues. Thus, these results strongly indicate that GR expression may contribute to GC sensitivity in canine MCT.     

Simple capillary electrophoretic determination of soluble oxalate and nitrate in forage grasses.

A simple capillary electrophoretic method is described for the simultaneous determination of soluble oxalate and nitrate in forage grasses. Grass samples were ground and extracted with water. The extracts were filtered and submitted to capillary electrophoresis. Electrophoresis was performed in a 75 microm x 50 cm fused silica capillary with 30 mM sodium sulfate containing an electroosmotic flow modifier under constant voltage at -8 kV. Separated oxalate and nitrate were detected with direct UV absorption at 214 nm. The present method can be used for routine monitoring of the concentration of soluble oxalate and nitrate in grasses.     

Encephalomyocarditis (EMC) virus-induced myocarditis by different virus variants and mouse strains.

The mode of occurrence of encephalomyocarditis (EMC) virus-induced myocarditis in mice was pathologically and virologically investigated using 2 virus variants (highly diabetogenic EMC-D and non-diabetogenic EMC-B) and 2 mouse strains (diabetes-susceptible BALB/c and diabetes-resistant C57BL/6). Mice were inoculated with 10(5) PFU/head of the virus intraperitoneally and observed up to 7 days post inoculation (7DPI). As compared with EMC-B-infected BALB/c and EMC-D-infected C57BL/6 mice, EMC-D-infected BALB/c mice developed marked myocarditis and exhibited a heart virus titer of more than 100 times above that of the others after 4DPI. Electron microscopically, small aggregations of virus-like particles, with 20-25 nm in diameter, were found in the cytoplasm of degenerated cardiomyocytes showing mitochondrial and myofibrillar degeneration in EMC-D-infected BALB/c mice.     

Effect of Dietary Inclusion of Blue Mussel Extract on Growth and Body Composition of Japanese Flounder Paralichthys olivaceus

Dietary inclusion of a water-soluble fraction of blue mussel Mytilus galloprovincialis was examined as a feeding stimulant for juvenile Japanese flounder Paralichthys olivaceus . The control diet mainly consisted of fish meal, potato starch, and pollack liver oil. Five, 10, and 20% (weight/weight) of the control diet was exchanged with aqueous extracts of blue mussel meat in experimental groups. Fish of about 10 g in initial body weight were fed each diet to satiation, twice daily, 6 d per wk for 6 wk at 20 C. The final body weight, weight gain, and feed efficiency of fish fed the diets containing blue mussel extracts were significantly higher than those of fish fed the control diet. However, these parameters were not different among experimental groups containing blue mussel extract independent of the inclusion level of extract. A similar trend was shown in protein efficiency ratio as fish fed the control diet had a significantly lower protein efficiency ratio than the other dietary groups. Compared to the control diet, higher plasma protein and lower triglyceride were found in fish fed the diets with the extract, while other blood constituents were relatively similar for the dietary groups tested. On the other hand, whole-body crude lipid content and lipid retention of fish fed the diets with the extract were generally significantly higher than those of fish fed the control diet. Whole body crude protein was identical regardless of the dietary composition; however, protein retention of fish showed a similar trend to lipid retention.     

Regulation of secondary follicle growth by theca cells and insulin-like growth factor 1

Ovaries contain follicles at various stages of development, including primordial, primary, secondary, antral and Graafian follicles. Although the growth of these follicles is controlled to maintain regular ovulation, the mechanism through which this occurs remains unclear. In our study, we found that the growth rate of cultured secondary follicles separated from mice ovaries differed between follicles. After 4 days of culture, the size of some secondary follicles was markedly increased, while that of others had either slightly increased, remained unchanged or shrunk. We compared the expression levels of growth factors between these secondary follicles and found that the growth rate of cultured secondary follicles correlated with the expression level of insulin-like growth factor 1 (Igf1) mRNA. Igf1 mRNA expression level in secondary follicles containing theca cells was higher than that in secondary follicles without theca cells, and the granulosa cell proliferation around follicles containing theca cells was increased. Furthermore, an IGF1 inhibitor also inhibited the granulosa cell proliferation, and administration of IGF1 to secondary follicles without growth promoted granulosa cell proliferation. These results indicated that the theca cells of secondary follicles induced the expression of IGF1 and promoted the follicle growth.     

Decomposition of the modulated waves in a rotating couette system

The time-dependent velocity field in a rotating Couette system has been decomposed. Successive instantaneous velocity profiles were obtained by an ultrasonic Doppler velocity profile method, and the spatiotemporal velocity field had been analyzed by two-dimensional Fourier transform. The Fourier spectrum is depicted by intrinsic peaks, and the velocity field cpan be reconstructed with these peak components only, indicating a successful decomposition of the velocity field. This method shows that the so-called broadband component, which is purely spatiotemporal and attributed to chaos, corresponds to a global motion of the wave propagating from roll to roll.     

Monoacylglycerol from Punica granatum seed oil.

The seeds of Punica granatum, known as hap roman in the Arabian Peninsula, are commonly eaten as a dessert. As part of an ongoing project to find nonnutritional natural products which have health benefits, or that can be exploited to protect crops, the chloroform-soluble extract of the fermented seeds of P. granatum was found to be rich in 1-O-trans,cis,trans-9,11,13-octadecatrienoyl glycerol (1). The seed oil is not lethal to brine shrimp larvae. 1-O-isopentyl-3-O-octadec-2-enoyl glycerol (2) and the known cis-9-octadecenoic, octadecanoic, and eicosanoic acids were also detected in small amounts in the seed oil by LC and MS. The structure of 1 was determined from NMR and MS spectral data.     

Antioxidant mechanism of carnosic acid: structural identification of two oxidation products.

To determine the antioxidant mechanism of food phenolics against the oxidation of food components, the reaction of carnosic acid, an antioxidative constituent of the popular herbs sage and rosemary, was investigated in the presence of ethyl linoleate and the radical oxidation initiator 2,2'-azobis(2,4-dimethylvaleronitrile). During this process, carnosic acid was oxidized to an o-quinone and a hydroxy p-quinone, the chemical structures of which were confirmed by physical and chemical techniques. From a quantitative time course analysis of the production of these quinones, an antioxidant mechanism of carnosic acid is proposed, consisting of the oxidative coupling reaction with the peroxyl radical at the 12- or 14-position of carnosic acid and subsequent degradation reactions.     

Recovery mechanism of the antioxidant activity from carnosic acid quinone,an oxidized sage and rosemary antioxidant

A solution of carnosic acid quinone, which is a radical chain-termination product having no antioxidant activity in the antioxidant reaction of carnosic acid, recovers potent antioxidant activity upon standing. The HPLC analysis of an aged solution of carnosic acid quinone revealed that several antioxidants are produced in the solution. From the time-course and quantitative analyses of the formation of the products and their structural analysis, an antioxidant mechanism from carnosic acid quinone is proposed that includes a redox reaction of carnosic acid quinone in addition to the isomerization to lactone derivatives. In the first stage of antioxidation, carnosic acid, the reduction product from carnosic acid quinone, contributes to the potent antioxidant activity of the solution. This proposed mechanism can explain one of the reasons for the strong antioxidant activity of the extract of the popular herbs sage and rosemary.     

Studies on the bacteria isolated anaerobically from paddy field soil III. Production of fatty acids and ammonia by Clostridium species

The volatile fatty acids produced by the representative strains of clostridia isolated from paddy field soil were surveyed. All the strains, except Cl. tertium, utilized only amino acids as the sole source of energy for growth. All of them produced acetic and butyric acids, and some of them produced propionic acid in addition. Iso-valeric and iso-butyric acids were also produced, except by Cl. tertium. These results suggest that all the strains, except Cl. tertium, may carry out the STICKLAND reaction. The cells of each strain harvested from the culture in the VL medium were subjected to examination whether or not they carried out the STICKLAND reaction in the presence of leucine or glycine. All the strains, except Cl. tertium, catalyzed the STICKLAND reaction and some strains also metabolized leucine and glycine even when they were supplied singly.